| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1386-1 | ||||
Resumo:Microbiome manipulation experiments involving corals and sponges as holobionts are challenging, mainly concerning variability of their microbiomes-associated in response to different environmental factors and stressors agents. To address this issue, the development of experimental systems capable of maintaining stable physicochemical conditions is essential, especially those used to simulate cold-water deep-sea (CWDS) ecosystems. A particular challenge arises when rearing CWDS corals and sponges since they are acclimated to a stable environment with absence of light, low nutrient levels and reduced temperature variation (5.5˚C ± 2˚C). Additionally, their microbiome has exhibited plasticity, indicating its ability to adapt to changing conditions. This characteristic introduces an additional confounding factor in microbiome manipulation experiments. Our aim in this study was to develop a system able to perform microbiome studies in CWDS coral and sponges. The Deep Sea Simulator (DSS) was a pioneering system developed for the PROBIODEEP project, designed in 3D SketchUp software and it consists of five essential sections distributed in a 130,000 L reservoir that efficiently pumps water to a temperature-controlled chemostat room, a water treatment system equipped with ozone-UV sterilization to prevent microorganisms entrance, a pre-treatment area responsible for controlling parameters such as pH, temperature, Oxidation reduction potential (ORP) and salinity through a controller, a cold chamber with stand-mounted aquaria, crucial for the CWDS corals and sponges cultivation and a mesocosm for conducting microbiome manipulation experiments. The enriched chemostat water with microorganisms flowed by gravity into the coral and deep-sea sponge cultivation system, as well as for the deep-sea mesocosm, composed of four replicating water baths and up to 40 two liters aquaria. The water flow rate was regulated using a 500 L/h industrial peristaltic pump to ensure continuous microbial culture growth. The ozone-UV water treatment system was effective to avoid cell invasion in the chemostat evaluated by flow cytometry cell count (96.41%). The DSS allowed the coral Desmophillum pertusa, the sponges Calyx sp., Aphrocallistes beatrix and Geodia sp. and their bacterial communities cultivation for at least 24 months and enabled a microbiome manipulation assay with increased temperature. These specimens were collected using a Remotely Operated Vehicle (ROV) at 600-800m depth. In summary, the DSS has proved to be a valuable tool for advanced microbiome research, offering stable and controllable biotic and abiotic conditions. Its design enables the reproducibility and repeatability of experimental conditions, thereby paving the way for comprehensive microbiome studies in CWDS corals and sponges. This innovation represents a significant leap forward in our ability to comprehend and manipulate these crucial ecosystems, providing insights into microbiome studies of CWDS corals and sponges. Palavras-chave: Microbiome, Mesocosm, cold water corals , cold water sponges, deep-sea Agência de fomento:Shell Brasil Petróleo LTDA | |||||